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Production of recombinant mink growth hormone in E. coli

Abstract

Escherichia coli cells expressing mink (Mustela vison) growth hormone were grown in a batch fermentation process. The expression level was estimated to be 27% of the total cellular protein after 3 h of induction with 1 mM isopropyl β-d-thiogalactoside (IPTG). If the expression of mink growth hormone (mGH) was induced with 0.2 mM IPTG, the concentration of target protein was slightly lower and was found to be 23% at the same time after induction. mGH expressed as inclusion bodies was solubilized in 8 M urea and renatured by dilution protocol at a protein concentration of 1.4–2.1 mg/ml in the presence of glutathione pair in a final concentration of 11.3 mM. [GSH]/[GSSG] ratio equal to 2/1 was used. Two-step purification process comprising of ion-exchange chromatography on Q-Sepharose and hydrophobic chromatography on Phenyl-Sepharose was developed. Some 25–30 mg of highly purified and biologically active mGH was obtained from 4 g of biomass. The method presented in this study allows producing large quantities of mGH and considering initiation of scientific investigation on mGH effect on mink in vivo and availability in fur industry.

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Correspondence to Jolanta Sereikaite.

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Sereikaite, J., Statkute, A., Morkunas, M. et al. Production of recombinant mink growth hormone in E. coli . Appl Microbiol Biotechnol 74, 316–323 (2007). https://doi.org/10.1007/s00253-006-0673-2

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Keywords

  • Growth Hormone
  • Inclusion Body
  • Nonreducing Condition
  • Total Cellular Protein
  • Inclusion Body Formation